Campylobacter assessment along the Spanish food chain: Identification of key points.

AIMS: Campylobacteriosis, caused by Campylobacter spp., is one of the most important foodborne zoonotic diseases in the world and a common cause of gastroenteritis. In the European Union, campylobacteriosis is considered the most common zoonotic disease, with over 10,000 cases in 2020 alone. This high occurrence highlights the need of more efficient surveillance methods and identification of key points. METHODS AND RESULTS: Herein, we evaluated and identified key points of Campylobacter spp. occurrence along the Spanish food chain during 2015-2020, based on the following variables: product, stage and region. We analysed a dataset provided by the Spanish Agency for Food Safety and Nutrition using a machine learning algorithm (random forests). Campylobacter presence was influenced by the three selected explanatory variables, especially by product, followed by region and stage. Among the studied products, meat, especially poultry and sheep, presented the highest probability of occurrence of Campylobacter, where the bacterium was present in the initial, intermediate and final stages (e.g., wholesale, retail) of the food chain. The presence in final stages may represent direct consumer exposure to the bacteria. CONCLUSSIONS: By using the random forest method, this study contributes to the identification of Campylobacter key points and the evaluation of control efforts in the Spanish food chain.

Spatial and temporal analysis of African swine fever front-wave velocity in wild boar: implications for surveillance and control strategies.

The front-wave velocity of African swine fever (ASF) virus spread is depicted through a retrospective spatial and temporal analyses of wild boar outbreaks from Jan. 2014 to Jan. 2022 in Estonia, Latvia, Lithuania and Eastern Poland-regions responsible for more than 50% of all wild boar cases in the EU. The study uses empirical semivariograms in a universal kriging model to assess spatial autocorrelation in notification dates and identifies a discernable large-scale spatial trend. The critical parameter of ASF front-wave velocity was identified (Mean = 66.33 km/month, SD = 163.24) in the whole study area, and explored the variations across countries, wild boar habitat suitability, seasons, and the study period. Statistical differences in front-wave velocity values among countries and temporal clusters are explored, shedding light on potential factors influencing ASF transmission dynamics. The implications of these findings for surveillance and control strategies are discussed.

First detection of herpesvirus and hemosporidians in the endangered Pyrenean Capercaillie (Tetrao urogallus aquitanicus).

Pathogens affect wild bird populations worldwide, contributing to their decline. Considering the scarce health data regarding the endangered Pyrenean Capercaillie (Tetrao urogallus aquitanicus), we molecularly surveyed selected pathogens (Newcastle disease virus, Avian influenza virus, Chlamydia psittaci, avian pathogenic Escherichia coli, Campylobacter jejuni, and Salmonella spp.) in 30 Pyrenean Capercaillie feces collected in the field (Catalonia, northeastern Spain). Additionally, swab and tissue samples from eight wild Pyrenean Capercaillies of Catalonia and Andorra were molecularly tested for herpesvirus and hemosporidians (Plasmodium spp., Haemoproteus spp., and Leucocytozoon spp.). All fecal samples were negative for the pathogens tested. Nevertheless, we detected a novel herpesvirus in 50% (4/8) of the Pyrenean Capercaillies, and hemosporidian DNA in 62.5% (5/8) of the tissue samples (i.e., Haemoproteus sp. [4 of 8] and/or Leucocytozoon sp. [3 of 8]). To our knowledge, this is the first detection of herpesvirus and hemosporidians infections in Pyrenean Capercaillies. The putative novel herpesvirus belongs to the genus Iltovirus. The presence of hemosporidian parasites in this mountain bird species is of concern, and could be related to the marked increase in the average temperature in the Pyrenees as a consequence of climate change. Our findings are fundamental to improve the conservation plans for the endangered Pyrenean Capercaillie population.

Salmonella assessment along the Spanish food chain: Likelihood of Salmonella occurrence in poultry and pig products is maintained across the food chain stages.

Salmonellosis is one of the most important foodborne diseases worldwide, including the European Union. Despite the One Health approach measures for risk assessment and risk management implemented by the European Union, the occurrence of disease and disease outbreaks remains high (e.g. 694 outbreaks were reported in 2020), highlighting the need of new assessment methods. Herein we applied machine learning using the random forests method to evaluate and identify key points regarding the occurrence of Salmonella sp. along the Spanish food chain during 2015-2020, using data provided by the Spanish Agency for Food Safety and Nutrition. We compared the role of the three categorical variables [product (20 categories), region (18 categories) and stage (11 categories)]. Salmonella presence was influenced by the three explanatory variables considered: first by product, followed by region and stage. The most determinant product for Salmonella probability was 'meat', while the most important stage was 'slaughterhouse'. Specifically, the highest values were found in pig and poultry meats. In these products, the Salmonella probability was high at the early and final stages of the food chain, although not at intermediate stages. The presence of Salmonella in the final stages (retail) of the food chain is of concern, as it can cause human cases of salmonellosis, including outbreaks. This study demonstrates the utility of the random forest method to identify key points and evaluate the control efforts. We recommend improving the surveillance and control measures, especially in the product and stages pointed out by our analysis, and enhancing the data collection harmonization among the different autonomous communities.

Analysis of Specific Physical Fitness in High-Level Table Tennis Players-Sex Differences.

Table tennis performance depends on multiple factors such as technique, tactics and fitness. Several studies have focused on investigating different technical-tactical variables. However, research analysing the specific physical qualities of this sport is scarce, particularly in the female sex. The aim of the present study was to assess the physical fitness variables associated with individual performance in elite table tennis players according to sex. Forty-eight elite players divided into males (n = 24; 25.38 ± 4.01 years) and females (n = 24; 22.33 ± 3.83 years) participated in the study. To determine physical fitness, participants performed vertical jump, hand grip strength, ergospirometry and lateral displacement tests (reaction time, displacement time and lateral acceleration). Male players showed higher values in vertical jump, hand grip strength and maximum oxygen consumption (p < 0.001). Likewise, male players moved laterally faster (p < 0.001). On the other hand, female players had a better reaction time towards the dominant side (p < 0.01). Elite male table tennis players showed better physical fitness compared to female players. Due to the scarcity of data on elite table tennis players, these results can serve as reference values for different table tennis practitioners.

Identifying the spatiotemporal vulnerability of soils to antimicrobial contamination through land application of animal manure in Minnesota, United States.

Antimicrobials may reach the soil environment from a variety of sources and pathways, including land application of human biosolids and animal manure. Once in soil, antimicrobials can affect the abundance and activity of soil microorganisms and exert selection pressures that enhance the emergence and spread of antimicrobial resistance (AMR). To mitigate the spread of AMR it is important to understand the spatial and temporal interactions between antimicrobials and soil. The goal of this study was to assess the vulnerability of Minnesota (U.S.) soil to contamination with specific antimicrobial compounds at temperatures experienced throughout the year. Soil contamination potential was estimated based upon specific antimicrobial drug binding and permanence, and average monthly temperature. Minnesota soil vulnerability was estimated by incorporating spatially explicit soil contamination potential, land cover type, and livestock density. Assessment of antimicrobials used in livestock production showed that soils are most vulnerable to antimicrobial contamination in southwestern Minnesota, to enrofloxacin, chlortetracycline, and oxytetracycline, and in the months of April and October. While the assessment herein was not based on actual on-farm antimicrobial use data and subsequent excretion of antimicrobial metabolites into the environment, this study provides an overview of the spatial and temporal potential for Minnesota soil to be contaminated by several antimicrobial drugs and demonstrates how specific vulnerability assessments might be conducted for geographic areas with known exposure (e.g., cropland fertilized with livestock manure and/or human biosolids). Such assessments might be used to identify best practices for mitigating antimicrobial exposure to soils and guide additional research to understand the role of environmental antimicrobial contamination in the problem of AMR.

African Swine Fever Survey in a European Context.

African swine fever (ASF) is currently the most threatening disease for domestic and wild pigs worldwide. Wild boar has been the main affected species in all EU countries except for Romania, where most notifications occur in domestic pigs. The spread of ASF in wild boar is challenging to control; risk factors are harder to identify and establish than in domestic pigs, which, together with an underestimation of the disease and the lack of treatment or an effective vaccine, are hindering control and eradication efforts. We distributed two online questionnaires, one for domestic pigs and one for wild boar, to experts of different background and countries in Europe, to explore risk factors in relation to ASF control connected to farming, hunting, trade, the environment, and domestic pig and wild boar populations. Overall, wild boar movements were estimated to pose the highest risk of ASF introduction and spread. The movement of pork and pork products for own consumption also ranked high. Here we explored, in addition to the assessment of risk pathways, the identification of risks of transmission at the domestic/wild boar interface, the importance of biosecurity practices and improved control efforts, and controversial opinions that require further attention.

Phytotoxic Effects of Antibiotics on Terrestrial Crop Plants and Wild Plants: A Systematic Review.

This review examines the state of knowledge on the phytotoxic effects of antibiotics on terrestrial crop plants and wild (non-crop) plants with the goal of evaluating differences in their sensitivity. This is important because environmental risk assessments of antibiotics currently consider their potential effects only on crop species but not wild species. Overall, we analysed 275 datasets consisting of antibiotic-plant species-endpoint combinations for germination (mg/L) and 169 datasets for plant growth (elongation and biomass) (mg/kg). EC10 and EC50 of each parameter were compared using a quotient approach, in which the geometric mean and the 5th percentile of the crop data were divided by wild data. Quotients were > 1 for elongation growth, suggesting that wild species were more sensitive than crops, while they were < 1 for biomass growth, suggesting quite the contrary. However, < 1% of the data in each dataset came from wild species, preventing definitive conclusions. Merging crop and wild data to evaluate differences in sensitivity among classes of antibiotics and plant families, we found using a linear mixed effect model and post hoc test that plants were most sensitive to phenicol and least sensitive to macrolides and tetracyclines. Further work must be conducted to gain a better understanding of the phytotoxic effects of antibiotics on terrestrial wild plants and subsequently assess whether the current approach to environmental risk assessment of antibiotics is sufficient to protect plant biodiversity.

Effects of a Novel Infant Formula on Weight Gain, Body Composition, Safety and Tolerability to Infants: The INNOVA 2020 Study.

Exclusive breastfeeding is recommended for the first six months of life to promote adequate infant growth and development, and to reduce infant morbidity and mortality. However, whenever some mothers are not able to breastfeed their infants, infant formulas mimicking human milk are needed, and the safety and efficacy of each formula should be tested. Here, we report the results of a multicenter, randomized, blinded, controlled clinical trial that aimed to evaluate a novel starting formula on weight gain and body composition of infants up to 6 and 12 months, as well as safety and tolerability. For the intervention period, infants were divided into three groups: group 1 received formula 1 (Nutribén® Innova 1 (Alter Farmacia S.A., Madrid, Spain) or INN (n = 70)), with a lower amount of protein, a lower casein to whey protein ratio by increasing the content of α-lactalbumin, and a double amount of docosahexaenoic acid/arachidonic acid than the standard formula; it also contained a thermally inactivated postbiotic (Bifidobacterium animalis subsp. lactis, BPL1TM HT). Group 2 received the standard formula or formula 2 (Nutriben® Natal (Alter Farmacia S.A., Madrid, Spain) or STD (n = 70)) and the third group was exclusively breastfed for exploratory analysis and used as a reference (BFD group (n = 70)). During the study, visits were made at 21 days and 2, 4, 6, and 12 months of age. Weight gain was higher in both formula groups than in the BFD group at 6 and 12 months, whereas no differences were found between STD and INN groups either at 6 or at 12 months. Likewise, body mass index was higher in infants fed the two formulas compared with the BFD group. Regarding body composition, length, head circumference and tricipital/subscapular skinfolds were alike between groups. The INN formula was considered safe as weight gain and body composition were within the normal limits, according to WHO standards. The BFD group exhibited more liquid consistency in the stools compared to both formula groups. All groups showed similar digestive tolerance and infant behavior. However, a higher frequency of gastrointestinal symptoms was reported by the STD formula group (n = 291), followed by the INN formula (n = 282), and the BFD groups (n = 227). There were fewer respiratory, thoracic, and mediastinal disorders among BFD children. Additionally, infants receiving the INN formula experienced significantly fewer general disorders and disturbances than those receiving the STD formula. Indeed, atopic dermatitis, bronchitis, and bronchiolitis were significantly more prevalent among infants who were fed the STD formula compared to those fed the INN formula or breastfed. To evaluate whether there were significant differences between formula treatments, beyond growth parameters, it would seem necessary to examine more precise health biomarkers and to carry out long-term longitudinal studies.

Physiological Profile, Metabolic Response and Temporal Structure in Elite Individual Table Tennis: Differences According to Gender.

No research that has analyzed the structural characteristics, physiological profile, and energy demands in the game of table tennis as played by women is available. The present study aimed to evaluate the physiological, metabolic, and temporal variables of table tennis players and to observe gender differences. Forty-eight elite table tennis players participated in this study: 24 men (25.3 ± 4.07 years) and 24 women (22.3 ± 3.8 years). During simulated competition, temporal structure, heart rate (HR), and lactate (LA) were evaluated. The maximum ergospirometric evaluations were performed in a laboratory. The total table tennis (TT) time and the total resting time (TRT) were longer for men (p < 0.05), but game density was higher for women (p < 0.05). During rallies, the real playing time (RPT) was longer for women, while the TRT was longer for men (p < 0.05). The maximum HR, minimum HR, and maximum LA concentrations were higher for men (p < 0.05). The obtained data reveal gender differences in the physiological, metabolic, structural, and temporal variables in table tennis players. The analysis of the studied variables could allow training sessions to be planned and organized according to table tennis players' gender.

Corrigendum: Evolution of the ASF Infection Stage in Wild Boar Within the EU (2014-2018).

[This corrects the article DOI: 10.3389/fvets.2020.00155.].

Spatial Trends in Salmonella Infection in Pigs in Spain.

Salmonella is one of the most important foodborne pathogens worldwide. Its main reservoirs are poultry and pigs, in which infection is endemic in many countries. Spain has one of the largest pig populations in the world. Even though Salmonella infection is commonly detected in pig farms, its spatial distribution at the national level is poorly understood. Here we aimed to report the spatial distribution of Salmonella-positive pig farms in Spain and investigate the presence of potential spatial trends over a 17-year period. For this, data on samples from pigs tested for Salmonella in 2002-2013, 2015, 2017, and 2019 as part of the Spanish Veterinary Antimicrobial Resistance Surveillance program, representing 3,730 farms were analyzed. The spatial distribution and clustering of Salmonella-positive pig farms at the province level were explored using spatial empirical Bayesian smoothing and global Moran's I, local Moran's I, and the Poisson model of the spatial scan statistics. Bayesian spatial regression using a reparameterized Besag-York-Mollié Poisson model (BYM2 model) was then performed to quantify the presence of spatially structured and unstructured effects while accounting for the effect of potential risk factors for Salmonella infection at the province level. The overall proportion of Salmonella-positive farms was 37.8% (95% confidence interval: 36.2-39.4). Clusters of positive farms were detected in the East and Northeast of Spain. The Bayesian spatial regression revealed a West-to-East increase in the risk of Salmonella infection at the province level, with 65.2% (50% highest density interval: 70-100.0%) of this spatial pattern being explained by the spatially structured component. Our results demonstrate the existence of a spatial variation in the risk of Salmonella infection in pig farms at the province level in Spain. This information can help to optimize risk-based Salmonella surveillance programs in Spain, although further research to identify farm-level factors explaining this pattern are needed.

Assessing the benefits of composting poultry manure in reducing antimicrobial residues, pathogenic bacteria, and antimicrobial resistance genes: a field-scale study.

The poultry industry in the European Union produces 13 million tons of manure annually, which represents a major health and environmental challenge. Composting is an environmental-friendly technique for the management of manure, but there are few studies about antibiotic residues and antimicrobial resistances at a field scale. The goal of this study was to determine if the composting of poultry manure at a field scale would result in the reduction of antibiotic residues, pathogenic bacteria, and antibiotic resistance genes (ARGs) in the final fertilizer product. A 10-week composting of poultry manure spiked with enrofloxacin, doxycycline, and ciprofloxacin was performed. The determination of antibiotics residues and 22 selected ARGs was carried out together with the identification of bacteria by metagenomics. In the case of ciprofloxacin and doxycycline, a 90% decrease was observed after composting for 3 weeks. Sixteen ARGs were detected at the beginning of the experiment; 12 of them decreased from week 0 to week 10 (reduction of 73.7-99.99%). The presence of potentially pathogenic bacteria, such as, Campylobacter coli or commensal bacteria such as Escherichia coli decreases along the composting process. In conclusion, 10-week composting of poultry manure promotes the reduction of antibiotic residues and most of the ARGs and pathogenic bacteria.

Evolution of the ASF Infection Stage in Wild Boar Within the EU (2014-2018).

African swine fever (ASF) is one of the most important emerging transboundary diseases of pigs, causing trade restrictions, and a health impact on susceptible pigs. Nine countries in the continental European Union (Estonia, Lithuania, Latvia, Poland, Czech Republic, Bulgaria, Belgium, Romania, and Hungary) have been affected by ASF from 2014 to 2018 and it keeps spreading despite the efforts to control it. For a number of years, we have witnessed high case-fatality rates in wild boar found dead particularly in new infected areas, which is typical of the peracute and acute forms of the infection at the beginning of an ASF epidemic. Experimental evidence with currently circulating strains indicates that some infected animals can remain asymptomatic and might even survive the infection. An increased presence of virus of moderate virulence can complicate ASF diagnosis as well as the mitigation and control of the disease. We analyze the ASF surveillance data in wild boar in the four EU countries where ASF has been present for longer, comparing the spatial density of antibody positive notifications with the time ASF has been present per region. Results indicate an increasing annual distribution of notifications based on antibodies over nucleic acid detection in hunted wild boar in Estonia, Latvia and Poland. Potentially, Lithuania, and Poland seem to have experienced more acute forms in 2017 and 2018 than Latvia and Estonia. Overall there was a positive statistical correlation between time with infection (TWI) and antibody positive density, with some variations in certain regions, particularly of Lithuania and Estonia. The increasing trend in potential survivors (hunted wild boar with confirmed PCR negative and antibody positive results) enhances the importance of surveillance design to sample and test shot wild boar. In conclusion, surveillance data based on ASFV detection by PCR and serology can be used to assess the status of the epidemic in wild boar.

Genetic characterization of Carnivore Parvoviruses in Spanish wildlife reveals domestic dog and cat-related sequences.

The impact of carnivore parvovirus infection on wild populations is not yet understood; disease signs are mainly developed in pups and assessing the health of litters in wild carnivores has big limitations. This study aims to shed light on the virus dynamics among wild carnivores thanks to the analysis of 213 samples collected between 1994 and 2013 in wild ecosystems from Spain. We determined the presence of carnivore parvovirus DNA by real-time PCR and sequenced the vp2 gen from 22 positive samples to characterize the strains and to perform phylogenetic analysis. The presence of carnivore parvovirus DNA was confirmed in 18% of the samples, with a higher prevalence detected in wolves (Canis lupus signatus, 70%). Fourteen sequences belonging to nine wolves, three Eurasian badgers (Meles meles), a common genet (Genetta genetta) and a European wildcat (Felis silvestris) were classified as canine parvovirus 2c (CPV-2c); five sequences from three wolves, a red fox (Vulpes vulpes) and a stone marten (Martes foina) as CPV-2b; and three sequences from a badger, a genet and a stone marten as feline parvovirus (FPV). This was the first report of a wildcat infected with a canine strain. Sequences described in this study were identical or very close related to others previously found in domestic carnivores from distant countries, suggesting that cross-species transmission takes place and that the parvovirus epidemiology in Spain, as elsewhere, could be influenced by global factors.

The Incorporation of Extracellular Vesicles from Mesenchymal Stromal Cells Into CD34+ Cells Increases Their Clonogenic Capacity and Bone Marrow Lodging Ability.

Mesenchymal stromal cells (MSC) may exert their functions by the release of extracellular vesicles (EV). Our aim was to analyze changes induced in CD34+ cells after the incorporation of MSC-EV. MSC-EV were characterized by flow cytometry (FC), Western blot, electron microscopy, and nanoparticle tracking analysis. EV incorporation into CD34+ cells was confirmed by FC and confocal microscopy, and then reverse transcription polymerase chain reaction and arrays were performed in modified CD34+ cells. Apoptosis and cell cycle were also evaluated by FC, phosphorylation of signal activator of transcription 5 (STAT5) by WES Simple, and clonal growth by clonogenic assays. Human engraftment was analyzed 4 weeks after CD34+ cell transplantation in nonobese diabetic/severe combined immunodeficient mice. Our results showed that MSC-EV incorporation induced a downregulation of proapoptotic genes, an overexpression of genes involved in colony formation, and an activation of the Janus kinase (JAK)-STAT pathway in CD34+ cells. A significant decrease in apoptosis and an increased CD44 expression were confirmed by FC, and increased levels of phospho-STAT5 were confirmed by WES Simple in CD34+ cells with MSC-EV. In addition, these cells displayed a higher colony-forming unit granulocyte/macrophage clonogenic potential. Finally, the in vivo bone marrow lodging ability of human CD34+ cells with MSC-EV was significantly increased in the injected femurs. In summary, the incorporation of MSC-EV induces genomic and functional changes in CD34+ cells, increasing their clonogenic capacity and their bone marrow lodging ability. Stem Cells 2019;37:1357-1368.

Salmonella Surveillance Systems in Swine and Humans in Spain: A Review.

Non-typhoid salmonellosis is a common and problematic foodborne zoonotic disease in which pork and pork products can be an important potential source of infection. To prevent this disease, important efforts to monitor the situation in the main source, livestock, are conducted in most developed countries. In the European Union, European Food Safety Agency (EFSA) and European Center for Disease Control (ECDC) compile information at the member-state level, even though important differences in production systems and surveillance systems exist. Here, Salmonella surveillance systems in one of the main sources of foodborne salmonellosis, swine, and humans in Spain were reviewed to identify potential gaps and discuss potential ways of integration under a "One-Health" approach. Despite the extensive information generated through the surveillance activities, source attribution can be only routinely performed through ad-hoc outbreak investigations, and national reports on human outbreaks do not provide sufficiently detailed information to gain a better understanding of the epidemiology of the pathogen. Human and animal monitoring of Salmonella would benefit from a better exchange of information and collaboration. Analysis of spatio-temporal trends in livestock and humans could help to identify likely sources of infection and to target surveillance efforts in areas with higher prevalence or where specific strains are found.